The EpiQuik™ Total Histone Extraction HT Kit provides a simple and high throughput microplate method for extracting histone proteins used for a variety of applications, which include histone modifications such as acetylation, methylation, and sumoylation. This kit is also specifically designed to meet the requirements of histone extracts used in EpiQuik™ histone quantification assays. The kit can be used to directly extract histones from mammalian cells cultured in a 96-well plate. The EpiQuik™ Total Histone Extraction HT Kit has the fastest procedure available on the market, allowing completion within 60 minutes.
- Pre-optimized and simple 1 hour protocol.
- Conveniently includes all essential reagents to carry out a histone extraction.
- Standardized procedure for reproducible results.
- Extracts a high yield of total core histones from as little as 104 cells.
- Post-translational modifications are kept intact.
- Does not affect histone modification status or levels.
Histones are the chief protein components of chromatin in biology. They act as spools around which DNA winds, and also play a role in gene regulation. The core histones include H2A, H2B, H3, and H4. Histones undergo post-translational modifications, which alter their interaction with DNA and nuclear proteins. The H3 and H4 histones have long tails protruding from the nucleosome, which can be covalently modified at several places. Modifications of the tail include methylation, acetylation, phosphorylation, ubiquitination, sumoylation, citrullination, and ADP-ribosylation (H2A can also be modified). Combinations of modifications are thought to constitute a code, the so-called "histone code." Histone modifications act in diverse biological processes such as gene regulation, DNA repair and chromosome condensation (mitosis).
Principle & Procedure
The EpiQuik™ Total Histone Extraction HT Kit simply applies our proprietary histone isolation buffers to cells. After treatment with Pre-lysis, Lysis, and Balance buffers, the total histones are easily extracted for immediate use or storage at proper conditions.
The minimal amount of starting materials can be as low as 104 cells. For the best results, the cell number should be greater than 3x104 cells. A total of 2x96 standard extractions (use 3x104 cells) can be performed with this kit.
Fig. 1. Schematic procedure for using the EpiQuik™ Total Histone Extraction HT Kit.
Fig. 2. Histone extracts were prepared from MCF-7 cells (30,000 cells/well) using the EpiQuik Total Histone Extraction HT Kit and acetyl histone H3 was quantified using the EpiQuik Global Pan-Methyl Histone H3K9 Quantification Kit (Colorimetric) (Cat. #P-3036).